Analysis of gentian violet residue in tissue

Abstract

A process for the analysis of gentian violet in animal or poultry tissue, animal or poultry manure, animal or poultry feed, or eggs. The gentian violet is quantitatively determined.

What is claimed is: 
     
       1. A process for the analysis of gentian violet in a material which comprises: (a) finely particulating a known amount of material containing gentian violet, unless such material is already in a finely divided state;   (b) admixing a known amount of acidified ethanol with said finely particulated material and letting the resultant admixture stand, said acidified ethanol removing said gentian violet from said material;   (c) separating said acidified ethanol (b) from said admixture (b) by centrifugation;   (d) removing any spectrophotometrical interfering substance from said acidified ethanol (c) by passing said acidified ethanol (c) through a chromatograph column, containing a basic sorbent, the acid in said acidified ethanol (c) being neutralized;   (e) concentrating said neutralized ethanol (d) so that the concentration of said gentian violet therein is increased;   (f) determining the amount of getian violet in said neutralized ethanol (e) and the background measurement of said neutralized ethanol (e) by spectrophotometrical means;   (g) quenching said gentian violet in said neutralized ethanol (e) by adding to said neutralized ethanol (e) a chemical substance which selectively renders said gentian violet non-detectable by spectrophotometric means without disturbing the spectrophotometric background of said neutralized ethanol;   (h) determining the background measurement of said neutralized ethanol (e) by spectrophotometrical means; and   (i) calculating the quantitative amount of gentian violet in said material using the spectrophotometrical measurements of steps (f) and (h).   
     
     
       2. A process as described in claim 1 wherein said material containing gentian violet is animal tissue, poultry tissue, eggs, animal feed, poultry feed, animal manure or poultry manure. 
     
     
       3. A process as described in claim 1 wherein said acidified ethanol was acidified by HCl. 
     
     
       4. A process as described in claim 1 wherein said basic sorbent is Alumin-AR. 
     
     
       5. A process as described in claim 1 wherein said neutralized ethanol (e) is concentrated in step (d) is concentrated by a factor of at least tenfold. 
     
     
       6. A process as described in claim 1 wherein said spectrophotometric measurements in steps (f) and (h) are made at 590 mμ. 
     
     
       7. A process for checking the accuracy of a process for the analysis of gentian violet in a material which comprises: (a) finely particulating a known amount of a material known not to contain any gentian violet, unless such material is already in a finely divided state;   (b) adding a known amount of at least one aqueous solution containing a known amount of gentian violet to a known amount of said material known not to contain any gentian violet to form at least one spiked sample;   (c) admixing a known amount of acidified ethanol with said spiked sample and letting the resultant admixture stand, said acidified ethanol removing said gentian violet from said material; (d) d. separating said acidified ethanol (b) from said admixture (c) by centrifugation;   (e) removing any spectrophotometrical interfering substance from said acidified ethanol (d) by passing said acidified ethanol (d) through a chromatograph column containing a basic sorbent, the acid in said acidified ethanol (d) being neutralized;   (f) concentrating said neutralized ethanol (d) so that the concentration of said gentian violet therein is increased;   (g) determining the amount of gentian violet in said neutralized ethanol (f) and the background measurement of said neutralized ethanol (f) by spectrophotometrical means;   (h) quenching said gentian violet in said neutralized ethanol (f) by adding to said neutralized ethanol (f) a chemical substance which selectively renders said gentian violet non-detectable by spectrophotometric means without disturbing the spectrophotometric background of said acidified ethanol;   (i) determining the background measurement of said neutralized ethanol (f) by spectrophotometrical means; and   (j) calculating the quantitative amount of gentian violet in said spiked material using the spectrophotometrical measurements of steps (g) and (i), said amount (i) of gentian violet for said spiked sample giving the accuracy of the procedure for the analysis of gentian violet in said spiked material by direct comparison with the known amount of gentian violet placed in said spiked sample in step (b).   
     
     
       8. A process as described in claim 7 wherein at least two spiked samples are prepared in step (b), each of said spiked samples having a different amount of gentian violet. 
     
     
       9. A process as described in claim 7 wherein said material containing gentian violet is animal tissue, poultry tissue, eggs, animal feed, poultry feed, animal manure or poultry manure. 
     
     
       10. A process as described in claim 7 wherein said basic sorbent is Alumin-AR. 
     
     
       11. A process as described in claim 7 wherein said neutralized ethanol is concentrated by a factor of at least tenfold. 
     
     
       12. A process as described in claim 7 wherein said acidified ethanol was acidified by HCl. 
     
     
       13. A process as described in claim 7 wherein said spectrophotometric measurements in steps (e) and (g) are made at 590 mμ. 
     
     
       14. A process for the analysis of gentian violet in a material which comprises: (a) finely particulating a known amount of material containing gentian violet, unless such material is already in a finely divided state;   (b) admixing a known amount of acidified ethanol with said finely particulated material and letting the resultant admixture stand, said acidified ethanol removing said gentian violet from said material;   (c) separating said acidified ethanol (b) from said admixture (b) by centrifugation;   (d) removing any spectrophotometric interfering substance from said acidified ethanol (c) by passing said acidified ethanol (c) through a chromatograph column, containing a basic sorbent, the acid in said acidified ethanol (c) being neutralized;   (e) determining the amount of gentian violet in said neutralized ethanol (d) and the background measurement of said neutralized ethanol (d) by spectrophotometrical means;   (f) quenching said gentian violet in said neutralized ethanol (d) by adding to said neutralized ethanol (d) a chemical substance which selectively renders said gentian violet non-detectable by spectrophotometric means without disturbing the spectrophotometric background of said neutralized ethanol;   (g) determining the background measurement of said neutralized ethanol (d) by spectrophotometrical means; and   (h) calculating the quantitative amount of gentian violet in said material using the spectrophotometrical measurements of steps (e) and (g).   
     
     
       15. A process as described in claim 14 wherein said material containing gentian violet is animal tissue, poultry tissue, eggs, animal feed, poultry feed, animal manure or poultry manure. 
     
     
       16. A process as described in claim 14 wherein said acidified ethanol was acidified by HCl. 
     
     
       17. A process as described in claim 14 wherein said basic sorbent is Alumin-AR. 
     
     
       18. A process as described in claim 14 wherein said spectrophotometric measurements in steps (e) and (g) are made at 590 mμ. 
     
     
       19. A process for checking the accuracy of a process for the analysis of gentian violet in a material which comprises; (a) finely particulating a known amount of a material known not to contain any gentian violet, unless such material is already in a finely divided state;   (b) adding a known amount of at least one aqueous solution containing a known amount of gentian violet to a known amount of said material known not to contain any gentian violet to form at least one spiked sample;   (c) admixing a known amount of acidified ethanol with said spiked sample and letting the resultant admixture stand, said acidified ethanol removing said gentian violet from said material;   (d) separating said acidified ethanol (b) from said admixture (c) by centrifugation;   (e) removing any spectrophotometrical interfering substance from said acidified ethanol (d) by passing said acidified ethanol (d) through a chromatograph column containing a basic sorbent, the acid in said acidified ethanol (d) being neutralized;   (f) determining the amount of gentian violet in said neutralized ethanol (e) and the background measurement of said neutralized ethanol (e) by spectrophotometrical means;   (g) quenching said gentian violet in said neutralized ethanol (e) by adding to said neutralized ethanol (e) a chemical substance which selectively renders said gentian violet non-detectable by spectrophotometric means without disturbing the spectrophotometric background of said acidified ethanol;   (h) determining the background measurement of said neutralized ethanol (e) by spectrophotometrical means; and   (i) calculating the quantitative amount of gentian violet in said spiked material using the spectrophotometrical measurements of steps (f) and (h), said amount (h) of gentian violet for said spiked sample giving the accuracy of the procedure for the analysis of gentian violet in spiked material by direct comparison with the known amount of gentian violet placed in said spiked sample in step (b).   
     
     
       20. A process as described in claim 19 wherein at least two spiked samples are prepared in step (b), each of said spiked samples having a different amount of gentian violet. 
     
     
       21. A process as described in claim 19 wherein said material containing gentian violet is animal tissue, poultry tissue, eggs, animal feed, poultry feed, animal manure or poultry manure. 
     
     
       22. A process as described in claim 19 wherein said basic sorbent in Alumin-AR. 
     
     
       23. A process as described in claim 19 wherein said acidified ethanol was acidified by HCl. 
     
     
       24. A process as described in claim 19 wherein said spectrophotometric measurements in steps (e) and (f) are made at 590 mμ.